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Am J Physiol Cell Physiol 284: C1497-C1507, 2003. First published February 5, 2003; doi:10.1152/ajpcell.00355.2002
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Vol. 284, Issue 6, C1497-C1507, June 2003

Na,K-ATPase inhibition alters tight junction structure and permeability in human retinal pigment epithelial cells

Sigrid A. Rajasekaran1, Jane Hu2, Jegan Gopal1, Ron Gallemore2,3, Sergey Ryazantsev4, Dean Bok2,5, and Ayyappan K. Rajasekaran1

1 Department of Pathology and Laboratory Medicine, 2 Jules Stein Eye Institute, 4 Department of Biological Chemistry, 5 Department of Neurobiology and Brain Research Institute, University of California, Los Angeles 90095; and 3 Retina-Vitreous Associates Medical Group, Los Angeles, California 90278

Na,K-ATPase regulates a variety of transport functions in epithelial cells. In cultures of human retinal pigment epithelial (RPE) cells, inhibition of Na,K-ATPase by ouabain and K+ depletion decreased transepithelial electrical resistance (TER) and increased permeability of tight junctions to mannitol and inulin. Electrophysiological studies demonstrated that the decrease in TER was due to an increase in paracellular shunt conductance. At the light microscopy level, this increased permeability was not accompanied by changes in the localization of the tight junction proteins ZO-1, occludin, and claudin-3. At the ultrastructural level, increased tight junction permeability correlated with a decrease in tight junction membrane contact points. Decreased tight junction membrane contact points and increased tight junction permeability were reversible in K+-repletion experiments. Confocal microscopy revealed that in control cells, Na,K-ATPase was localized at both apical and basolateral plasma membranes. K+ depletion resulted in a large reduction of apical Na,K-ATPase, and after K+ repletion the apical Na,K-ATPase recovered to control levels. These results suggest a functional link exists between Na,K-ATPase and tight junction function in human RPE cells.

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